The increasing anti-bacterial drug resistance is one of the biggest challenges facing doctors around the globe, so finding alternative treatments is one of the ideal options to overcome this problem. The cruciferous family is one of the wealthiest plants worldwide because it contains the most important secondary metabolites, glucosinolates, known for their anti-microbial properties. The present study aimed to evaluate the anti-bacterial effect of glucosinolates (Sinigrin) against eight bacterial isolates (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Actinomyces, Proteus mirabilis and Streptococcus pneumoniae). The current study investigated six concentrations of pure Sinigrin (100, 300, 500, 700, 900, and 1100 µg/ml). The sensitivity of bacterial isolates to various antibiotics was tested by VITIK 2DensiCheck equipment. The anti-bacterial activity of Sinigrin was assessed using the agar diffusion method, and the microtiter plate method measured the minimal inhibitory concentration (MIC). The highest anti-bacterial effect of Sinigrin was observed against S. aureus, E. coli, and E. faecalis. The anti-bacterial activity started as lower as 100 µg/ml, while a moderate effect was seen against P. aeruginosa and K. pneumoniae at a concentration lower than 700 µg/ml. On the other hand, Sinigrin was not effective against Actinomyces, P. mirabilis, and S. pneumoniae. It can be concluded from the present study that Sinigrin has an anti-bacterial effect on some isolates of bacteria which suggests the possibility of using Sinigrin as alternative medicine in the future. Keywords: Anti-bacterial activity, Agar well diffusion, Glucosinolates, Minimum inhibition concentration and antibiotic susceptibility, Sinigrin.